Formulation of Oil-in-Water Cream from Mangosteen

E. Isabella, T. Pohan. / Atom Indonesia Vol. 39 No. 3 (2013) 136 – 144

Formulation of Oil-in-Water Cream from Mangosteen (Garcinia mangostana L.) Pericarp Extract Preserved by Gamma Irradiation

E. Isabella* and T. Pohan Faculty of Pharmacy, Pancasila University, Srengseng Sawah, Jagakarsa, Jakarta 12640, Indonesia

A R T I C L E I N F O A B S T R A C T

Article history:

Received 24 November 2013

Received in revised form 20 December 2013

Accepted 26 December 2013

Keywords:

Anti-aging cream

Mangosteen pericarp extract

Garcinia mangostana L.

Decontamination

Gamma irradiation

The aim of this research was to formulate mangosteen (Garcinia mangostana L.)

pericarp extract into oil-in-water (O/W) cream which stable and durable. In order to

improve the shelf life durability of the formula, the irradiation of formula using

gamma rays from cobalt-60 was done. The concentrations of extract were varied to

1, 2 and 3% and were irradiated by gamma rays with doses of 5, 7.5 and 10 kGy.

Physicochemical and microbiological properties of these formulas were carried out

for 90 days stored at 30 ± 2°C and 40 ± 2°C. It was found that both of

physicochemical and microbiological properties of the formulas were changed by

extract concentrations and after irradiation. The irradiated formulas with dose up to

7.5 kGy, which were stored at 30 ± 2°C and 40 ± 2°C, presented acceptable

physicochemical and microbiological stability for at least 90 days. Analysis by

TLC (Thin Layer Chromatography)-densitometry to study decomposition of

G. mangostana pericarp extract cream formulas was carried out 24 h after

preparation and 90 day after storage. The result of TLC-densitometry analysis

showed that G. mangostana pericarp extract in the cream formulas did not develope

significant decomposition after 90 days of storage. Decontamination dose for all

formulas was found to be about 7.5 kGy. At this condition, the bacteria and mold-

yeast have been killed, without reducing the antioxidant activity.

© 2013 Atom Indonesia . All rights reserved

INTRODUCTION

Skin is subject to deterioration through

dermatological disorders, environmental abuse, or

through the normal aging process (chronoaging),

which may be accelerated by exposure of skin to sun

(photoaging). Environmental abuse such as

ultraviolet radiation, air pollutants, chemicals, air

conditioning, central heating, alcohols, daily stress

and other environmental factors can result in the

generation of free radicals in the exposed skin which

can lead to premature aging of skin [1]. The visible

signs of aging skin are wrinkles, lines, sagging,

hyperpigmentation and age spots [2,3].

The effective treatment and prevention of skin

conditions due to natural or environmental aging

may be obtained through the application of cosmetic

compositions to the skin which comprises collagen

stimulators, phytoestrogens, vitamins, allantoin,

placental extract, antioxidants [2], some of which,

such as vitamins and antioxidants can be found in

many plants. One type of plants that has potential

antioxidant activity is mangosteen.

Corresponding author.

E-mail address: [email protected]

Mangosteen (Garcinia mangostana L.) from

family Clusiaceae, is a tropical fruit from Southeast

Asian. People have used the pericarp of

G. mangostana as a traditional medicine for the

treatment of abdominal pain, diarrhea, dysentery,

infected wound and chronic ulcer [4]. Moreover,

recent scientific studies reported that pericarp of

G. mangostana is a source of xanthones and other

bioactive subtances which possessed several

biological and pharmacological properties such as

antioxidant, antiproliferative, antihistamine, anti-

fungal, antibacterial and induction of apoptosis in

cancer cell line [5-7]. In addition, Ochiai et al.

[8] reported that G. mangostana pericarp extract

have bioactive subtances which inhibit matrix

metalloproteinases, that are well known to be zinc-

containing proteolytic enzymes that hydrolyze

extracellular matrix-forming protein including

collagen and to cause changes due to the aging

of the skin.

The skin treatment composition of this

research is preferably on oil-in-water (O/W) type

emulsion since this type of emulsion affords better

cosmetic feel to the product, light when used, easily

washed and leave no greasy feeling [3]. Topically

applied formulas must not contain microbials

Atom Indonesia Vol. 39 No. 3 (2013) 136 - 144

136


exceeding the permissible limits. According to

The National Agency of Drugs and Food Controls

of Indonesia (NADFC = BPOM) [9], the maximum

contamination limit of cosmetics for microbes and

fungal is 103 cfu/g, respectively. Cobalt-60 source,

which is commonly used for gamma irradiation, can

be used for sterilization/decontamination of cosmetic

raw materials and finished products. Aiming at the

reduction in microbiological content, the method

cannot make something radioactive or leave any

residual radioactive and also can be processed at

room temperature. Gamma irradiation can penetrate

the packaging materials and sealed packages

containing the final product, thus destroying the

existing microorganisms and extend the shelf life of

products [10,11]. The objective of this research was

to develop anti-aging cream from G. mangostana

crude extract and to extend shelf life of formulas by

gamma irradiation.

EXPERIMENTAL METHODS

Materials and equipments

Garcinia mangostana fruits were purchased

from a local plantation in Leuwiliang, Bogor,

West Java. The authentication of the plant material

was carried out at the Herbarium Bogoriensis –

Research Center for Biology, Indonesian Institute of

Sciences, Cibinong, West Java.

Stearic acid, cetyl alcohol (1-hexadecanol),

isopropyl myristate, glyceryl monostearate,

propylene glycol, triethanolamine, and perfume

were purchased from Tritunggal Arthamakmur;

butylated hydroxytoluene, methylparaben,

propylparaben, and distilled water were purchased

from Brataco. DPPH (1,1-diphenyl-2-

picrylhydrazyl) radical, 70% ethanol, cerium sulfate,

methanol, chloroform and GF254 silica plate were

used in this research.

Equipments used were gamma irradiator

“Karet Alam” with cobalt-60 source (activity:

120 kCi at 2012), vacuumized rotary evaporator

(Büchi), densitometer (CAMAG), UV light

cabinet (CAMAG), spectrophotometer UV-VIS

(Shimadzu UV 1800), laminar air flow, oven

(Memmert), viscometer (Brookfield RV),

waterbath (Memmert), analytical balance

(Mettler Toledo AB 204), microscope (Olympus

Optical), stirrer (Eurostar), centrifuge (Porta),

extensometer, pH meter (Meterohm type 620),

ultrasonicator, hot plate, chromatograph

chamber, incubator, objective glass, thermometer

(Assistant).

Crude extract formulation

The pericarp of G. mangostana fresh fruits

were chopped into small pieces, dried and extracted

with 70% ethanol for one weeks, four times, at room

temperature. The filtrates were pooled and

concentrated by a vacuumized rotary evaporator

at 40°C.

O/W cream from G. mangostana pericarp extract formulation

Stearic acid, cetyl alcohol, isopropyl

myristate, glyceryl monostearate and butylated

hydroxytoluene were mixed, molten under heating

and maintained at 70°C (dispersed phase). The other

ingredients were mixed, dissolved under heating

and maintained at 70°C (dispersing phase).

The dispersed phase was gradually added to the

dispersing phase at 70°C with sufficient stirring,

G. mangostana pericarp extract and perfume were

mixed into the base at 40°C. It was then emulsified

to homogeneity by stirring (15 min, 150 rpm.min-1).

Table 1. Composition (%w/w) of O/W cream from

G. mangostana pericarp extract.

Components Formula code

A B C

G. mangostana pericarp extract 1 2 3

stearic acid 5 5 5

cetyl alcohol 2.75 2.75 2.75

isopropyl myristate 10 10 10

glyceryl monostearate 3 3 3

propylene glycol 10 10 10

triethanolamine 1 1 1

butylated hydroxytoluene 0.05 0.05 0.05

methylparaben 0.15 0.15 0.15

propylparaben 0.05 0.05 0.05

perfume (musk) 1 1 1

distilled water 66 65 64

Irradiation process

Irradiation was performed at room

temperature using cobalt-60 source from Gamma

Irradiator “Karet Alam”. All samples were irradiated

at doses of 5, 7.5 and 10 kGy. The samples were

G. mangostana pericarp extract and cream formulas

(code: A, B, C).

Organoleptic properties, pH, DPPH radical

scavenging activities and TLC profiles were carried

out for extract samples, while physicochemical

properties (organoleptic, emulsion type, centrifuge,

spreadability, viscosity and flow properties, mean

particle size and pH), microbiological (total plate

count) properties and TLC densitometry profiles

were carried out for cream formulas at room

137


temperature (30 ± 2°C) and high temperature

(40 ± 2°C).

Unirradiated formulas were used as controls

to detect the changes resulting from the action of

ionizing radiation on the formulas investigated.

DPPH radical scavening activities

The decrease of the absorption at 517 nm

of the DPPH solution after addition of the

G. mangostana extract samples (unirradiated and

irradiated) were measured in a glass cuvette

(1 cm length). Each of an aliquot (1 mL) of 0.4 mM

methanolic DPPH solution was added to 10,

20, 30, 40 and 50 µL of 1000 mg/L methanolic

G. mangostana extract samples solution so that the

concentrations in the cuvette were 2, 4, 6, 8 and

10 mg/L. All the tests were performed in triplicate at

37°C (incubation). The absorption was monitored at

30 min after incubation. Absorbance values were

corrected for radical decay using blank solutions.

Reduction of DPPH was followed by monitoring the

decrease in absorbance at 517 nm until the reaction

reached a plateau. The inhibition concentration

(IC50) is the amount of antioxidant necessary to

decrease the initial DPPH by 50%. The lower the

IC50 is, the higher the antioxidant activity.

TLC profiles of G. mangostana pericarp extract

Unirradiated and irradiated extract solutions

[1% (w/v)] were prepared and analyzed by TLC

using chloroform - methanol (12:1) as eluent.

The spots were observed and marked under UV 254

nm light cabinet, then sprayed with 1% cerium

sulfate reagent in 10% sulfuric acid and dried on the

hot plate to form visible spots.

Physicochemical properties

All test were performed on unirradiated

and irradiated cream formulas at doses of 5, 7.5, and

10 kGy. The results were analyzed statistically:

(1) Organoleptic properties: formulas were evaluated

by their color, odor and homogeneity; (2) Emulsion

type: formulas were mixed with sudan III on

the objective glass and investigated by light

microscope; (3) Centrifuge: formulas were placed in

centrifuge tubes, vigorously shaken for 15 min at

2500 rpm.min-1

and observed whether there was

a separate section or not; (4) Spreadability: formulas

were measured using extensometer; (5) Viscosity

and flow properties: formulas were measured

by Brookfield rheometer (RV type); (6) Mean

particle size: particle sizes of formulas were

investigated by light microscope. Results were in

an average; (7) pH: formulas were measured

by pHmeter.

Microbiological properties

Number of total bacteria and total mold-yeast

of the cream formulas were determined by

plate count method according to SNI 01-2897-1992

[12]. A total of 10 g cream formulas were added

into 90 mL of buffer phosphate solution (pH 7.2),

thus obtained the dilution of 1:10, homogenized,

then gradually diluted to 10-2

, 10-3

, 10-4

, 10-5

and 10-6

. Each of a total of 1 mL of diluted sample

was poured into a sterile Petri dish, 15-20 mL

of nutrient agar medium was added, shaken

gently until evenly mixed sample, silenced

until frozen, then the Petri dish was reversed

and incubated at 30-35°C for 48 h. Similar

as described in the determination number of

total bacteria, each of a total of 1 mL of

diluted sample was poured into a sterile Petri

dish, 15-20 mL of potato dextrose agar medium

was added, shaken gently until evenly mixed

sample, silenced until frozen, then the Petri dish

was reversed and incubated at 20-24°C for

5 days. The number of total bacteria and

total mold-yeast were calculated using the following

equation:

( ) (1)

where N is total plate count (cfu/g); ΣC is total

colony of all counted Petri dish; p, q, r are total

counted Petri dish at first, second, third dilution;

d is first counted dilution.

TLC-densitometry

G. mangostana pericarp extract was separated

from cream formulas and identified by TLC-

densitometry. The results were compared between

unirradiated and irradiated formulas. Each of a total

of 100 mg of formula was dissolved in methanol

(10000 mg/L), then 20 µL of 10000 mg/L formula

solution was spotted on GF254 silica plate and

analyzed by TLC using chloroform - methanol

(12:1) as eluent. The spots were observed by

densitometer at 317 nm.

138


Statistical analysis

Physicochemical properties (spreadability,

viscosity, mean particle size, pH) and

microbiological (total plate count) test results were

given as the mean of 3 experiments and ANOVA

two ways (p = 0.05) from Minitab 16 software

program was employed for analyses.

RESULTS AND DISCUSSION Mangosteen pericarp extract

Extraction of 4.5 kg of G. mangostana

pericarps, got 19.32% of crude extract.

Physicochemical properties of unirradiated and

irradiated G. mangostana pericarp extract were

summarized in Table 2. It was determined that there

was no change in the physicochemical properties of

irradiated G. mangostana pericarp extract at

3 different doses.

Table 2. Physicochemical properties and IC50 values of

G. mangostana pericarp extract before and after irradiation.

Properties Unirradiated Irradiated (kGy)

5 7.5 10

Organoleptic viscous, dark brown,

characteristic + + +

pH 5.45 5.55 5.55 5.60

IC50 (mg/L) 8.08 6.87 5.30 4.71 *(+ : no change; - : change)

The antioxidant activity of G. mangostana

pericarp extract which was implemented in linear

regression:

(2)

between extract concentration versus percent of

inhibition were shown in Fig. 1. The inhibition

concentration-fifty (IC50) which expresses the ability

of the extract to inhibit 50% of DPPH radical

calculated by substituting Y by 50 for each linear

regression curve, in order to obtain the IC50 values.

Fig. 1. Curve of extract concentration versus inhibition percentage.

Table 2 shows that the increase in radiation dose

until 10 kGy caused the decrease in the IC50 value,

which means that the antioxidant activity increased.

The extract is declared to have a very strong

antioxidant activity if the IC50 value < 50 mg/L

[13]. This fact suggested that the irradiation dose

until 10 kGy is still feasible for the microbial

decontamination of G. mangostana pericarp extract.

Based on the TLC profiles there were at least

3 spots observed in extract formulations (Fig. 2).

It was showed that the irradiated spots at the

3 different doses were not significantly different

from the unirradiated spots. The irradiation

dose until 10 kGy did not cause any significant

changes in TLC profiles of G. mangostana pericarp

extract.

Fig. 2. TLC of unirradiated and irradiated G. mangostana

pericarp extract (condition: silica gel GF254, chloroform -

methanol (12:1), UV light at 254 nm, 1% cerium sulphate in

10% sulfuric acid).

Physicochemical properties

Organoleptic properties, emulsion type and

centrifuge (physical properties) of unirradiated

and irradiated formulas were summarized in Table 3.

Physical properties of formulas stored at room

and high temperature did not suffer perceptible

alterations during the study period of 90 days.

The increasing extract concentrations and

irradiation doses affected its color parameter

unsignificantly.

Spreadability value of unirradiated and

irradiated formulas were summarized in Table 4.

It was determined that increasing extract

concentrations caused increasing spread ability

values, while increasing doses of irradiation caused

decreasing spread ability values as shown in Fig. 3.

Changes in spreadability value of all formulas

seem depends on extract concentrations (p<0.05)

and irradiation dose levels (p<0.05). Apparent

spreadability of formulas resulted in increased

values when stored at high temperature and during

90 days of storage.

(0 kGy)y=5,6295x+4,5433

(5 kGy)y=7,0765x+1,3637

(7.5 kGy)y=5,295x+21,964

(10 kGy)y=7,0602x+16,751

Extract concentration (mg/L)

Inhib

itio

n P

erc

enta

ge

90

80

70

60

50

40

30

20

10

0 2 4 6 8 10

139


Table 3. Physical properties of formulas before and after irradiation evaluated after 24 h of formulation and at 90th day of analyses.

C

o

d

e

Storage

conditions

24 h after formulation 90th day of analyses

Unirradiated Dose (kGy)


5 7.5 10 5 7.5 10 O ET C O ET C

A Room

temperature

30 ± 2°C

LB, M, H O/W x + + + LB, M, H O/W x + + +

B DB, M, H O/W x + + + DB, M, H O/W x + + +

C DB, M, H O/W x + + + DB, M, H O/W x + + +

A High

temperature

40 ± 2°C

LB, M, H O/W x + + + LB, M, H O/W x + + +

B DB, M, H O/W x + + + DB, M, H O/W x + + +

C DB, M, H O/W x + + + DB, M, H O/W x + + +

*O: organoleptic; LB: light brown; DB: dark brown; M: musk; H: homogen; ET: emulsion type; O/W: oil in water; C: centrifuge; x: no separation;

(+ : no change; - : change); n=3

Table 4. Spreadability value of formulas before and after irradiation evaluated after 24 h of formulation and at 90th day of analyses.

C

o

d

e

Storage

conditions




5 7.5 10 5 7.5 10

A

Room

temperature

30 ± 2°C

5787.91

± 34.61

5429.79

± 77.77

5061.85

± 53.83

4394.37

± 90.55

6351.52

± 56.11

6097.5

1 ± 34.83

5767.75

± 52.31

5091.43

± 82.46

B

6093.10

± 93.52

5763.30

± 63.92

5256.98

± 49.85

4616.14

± 46.01

6625.07

± 56.06

6325.69

± 67.67

5962.26

± 55.95

5308.47

± 38.92

C 6349.16

± 53.96

6056.18

± 72.41

5618.75

± 112.90

4891.20

± 90.36

7064.79

± 41.02

6627.48

± 57.87

6316.52

± 113.11

5523.69

± 59.83

A

High

temperature

40 ± 2°C

6062.99

± 44.88

5772.55

± 117.69

5519.51

± 101.50

4753.90

± 104.76

6685.31

± 58.52

6401.05

± 57.98

6095.27

± 59.60

5497.55

± 95.26

B 6375.05

± 46.03

6130.03

± 91.49

5774.49

± 57.27

5011.48

± 28.73

7284.97

± 58.69

6818.79

± 59.84

6377.40

± 38.96

5828.44

± 44.02

C 6968.39

± 68.66

6639.94

± 141.53

6169.66

± 132.92

5244.40

± 102.60

7722.40

± 66.15

7297.83

± 121.15

6943.70

± 48.39

6113.70

± 52.32

*values were represented as mean ± standard deviation, n=3

Fig. 3. Spreadability value of formulas before and after

irradiation (n=3).

Viscosity value of unirradiated and irradiated

formulas were summarized in Table 5. It was

determined that increasing extract concentrations

caused decreasing viscosity values, while increasing

doses of irradiation caused increasing viscosity

values as shown in Fig. 4. Changes in viscosity

value of all formulas seem depends on extract

concentrations (p<0.05) and irradiation dose levels

(p<0.05). Apparent viscosity of formulas resulted in

decreased values when stored at high temperature

and during 90 days of storage.

Fig. 4. Viscosity value of formulas before and after irradiation

(n=3).

0

1000

2000

3000

4000

5000

6000

7000

A B C

Spre

ad a

bili

ty (

mm

²)

0 kGy

5 kGy

7.5 kGy

10 kGy

80000

90000

100000

110000

120000

130000

A B C

Vis

cosity (

cP

s)

0 kGy

5 kGy

7.5 kGy

10 kGy

140


Table 5. Viscosity value of formulas before and after irradiation evaluated after 24 h of formulation and at 90th day of analyses.

C

o

d

e

Storage

conditions




5 7.5 10 5 7.5 10

A

Room

temperature

30 ± 2°C

105333.33

± 1755.94

107666.67

± 3253.20

117291.67

± 1572.88

124583.33

± 3145.76

94833.33

± 2254.63

100166.67

± 3253.20

105333.33

± 3253.20

115833.33

± 2602.08

B 94833.33

± 3013.86

96833.33

± 2753.79

102708.33

± 954.70

112083.33

± 1909.41

87333.33

± 2843.12

92000.00

± 3968.63

96333.33

± 1755.94

108958.33

± 3207.25

C 82666.67

± 2565.80

89333.33

± 1607.28

96250.00

± 4098.40

106666.67

± 3442.23

77666.67

± 2565.80

83833.33

± 3013.86

88666.67

± 1892.97

98500.00

± 2500.00

A

High

temperature

40 ± 2°C

79666.70

± 1892.97

84666.67

± 3253.20

90625.00

± 2724.31

102291.67

± 3145.76

70666.67

± 2565.80

76833.33

± 2843.12

82166.67

± 3752.78

95208.33

± 4434.12

B 67666.67

± 2565.80

78000.00

± 1322.88

79375.00

± 2864.11

97291.67

± 954.70

59666.67

± 2753.79

65666.67

± 3253.20

73166.67

± 2753.79

87916.67

± 2194.93

C 63833.33

± 1258.31

67833.33

± 1258.31

75416.67

± 2818.28

84583.30

± 3207.25

54333.33

± 2254.63

60333.33

± 1258.31

66000.00

± 3278.72

75333.33

± 3685.56


Table 6. Mean particle size value of formulas before and after irradiation evaluated after 24 h of formulation and at 90th day of

analyses.

C

o

d

e

Storage

conditions




5 7.5 10 5 7.5 10

A

Room

temperature

30 ± 2°C

81.46 ± 0.43 62.00

± 0.51

55.64

± 0.31

42.39

± 0.31 83.51 ± 0.37

65.35

± 0.51

59.41

± 0.42

47.98

± 0.65

B 83.13 ± 0.21 63.79

± 0.26

56.69

± 0.35

43.62

± 0.36 85.66 ± 0.37

67.67

± 0.31

61.61

± 0.34

49.35

± 0.34

C 84.43 ± 0.47 64.70

± 0.25

58.53

± 0.42

44.33

± 0.50 87.56 ± 0.29

69.82

± 0.33

63.73

± 0.26

52.57

± 0.45

A

High

temperature

40 ± 2°C

82.58 ± 0.37 63.95

± 0.44

56.85

± 0.29

45.31

± 0.26 87.53 ± 0.72

68.07

± 0.46

62.71

± 0.33

51.01

± 0.31

B 84.94 ± 0.37 65.58

± 0.43

58.98

± 0.32

46.34

± 0.36 90.44 ± 0.45 70.05

± 0.42

63.92

± 0.39

53.28

± 0.28

C 86.77 ± 0.38 67.25

± 0.39

60.68

± 0.35

47.42

± 0.28 91.71 ± 0.57 72.38

± 0.35

66.57

± 0.43

55.65

± 0.29


Fig. 5. Rheogram of formula A after irradiation dose of 7.5 kGy

at 90 days of analyses (n=3).

Figure 5 showed that formula A had thixotropic

plastic behaviour. These result represented that the

formula did not begin to flow until a shearing stress,

corresponding to the yield value, was exceeded.

At moderately high shear rate, formulation was easy

to apply to the skin.

Mean particle size value of unirradiated and

irradiated formulas were summarized in Table 6.

It was determined that increasing extract

concentrations caused increasing particle size

values, while increasing doses of irradiation caused

decreasing particle size values as shown at Fig. 6.

Changes in mean particle size value of all formulas


0

0,5

1

1,5

2

2,5

3

3,5

4

4,5

100000 200000 300000 400000

Shear

rate

(s-1

)

Shear stress (dyne/cm²)

Series1

Series2

Series3

141


and irradiation dose levels (p<0.05). Apparent mean

particle size of formulas resulted in increased values

when stored at high temperature and during 90 days

of storage.

Fig. 6. Mean particle size value of formulas before and after

irradiation (n=3).

The presence of G. mangostana pericarp extract at

the formulas affected their particle size values, due

to the increasing of water soluble components at the

dispersing phase from the bioactive subtances such

as tanin, flavonoid, alkaloid in G. mangostana

pericarp extract. Increasing of the dispersing phase

was not followed by emulgator enhancement in

formula, thus led to insufficient in an interfacial film

between the dispersed and dispersing phase,

the dispersing phase tended to form larger droplets,

that caused decreasing in viscosity values. The lower

the viscosity value is, the ability to spread is also

getting lower. At higher temperature condition

(40 ± 2°C), the particle size values had increased

due to the effect of the elevated temperature of

storage which caused an acceleration of the motion

rate of the dispersed phase.

pH value of unirradiated and irradiated

formulas were summarized in Table 7. It was

determined that increasing extract concentrations

and doses of irradiation caused decreasing pH

values, probably, due to the acidity of the extract, as

shown in Fig. 7. Changes in pH value of all formulas


and irradiation dose levels (p<0.05). The formulas

presented tendency to reduce the pH value at high

temperature and during 90 days of storage condition,

probably, due to the deterioration of the components

of the fluid emulsions and the bioactive substances

from the extract of G. mangostana.

Fig. 7. pH value of formulas before and after irradiation (n=3).

Microbiological properties

The microbial contamination value of

unirradiated and irradiated formulas were

summarized in Table 8. An irradiation dose of 5 kGy

on formulas could eliminate mold-yeast from

initial contamination while to remove bacterial

contamination completely, the higher dose is

needed. Bacterial contamination can be removed

completely by irradiation dose of ≥ 7.5 kGy.

Changes in microbial contamination of all formulas

seem not depends on extract concentrations (p>0.05)

and depends on irradiation dose levels (p<0.05).

High temperature storage condition did not affected

total microbial contamination value significantly

during 90 days of storage.

Table 7. PH value of formulas before and after irradiation evaluated after 24 h of formulation and at 90th day of analyses.

C

o

d

e

Storage

conditions




5 7.5 10 5 7.5 10

A Room

temperature

30 ± 2°C

6.62 ± 0.03 6.82 ± 0.03 6.88 ± 0.06 6.92 ± 0.03 6.35 ± 0.00 6.28 ± 0.03 6.27 ± 0.06 6.28 ± 0.03

B 6.33 ± 0.03 6.47 ± 0.03 6.63 ± 0.03 6.67 ± 0.03 6.17 ±0.03 6.12 ± 0.06 6.10 ± 0.05 6.02 ± 0.03

C 6.02 ± 0.03 6.12 ± 0.03 6.22 ± 0.03 6.28 ± 0.03 5.77 ± 0.06 5.68 ± 0.03 5.67 ± 0.06 5.67 ± 0.00

A High

temperature

40 ± 2°C

6.53 ± 0.03 6.67 ± 0.06 6.72 ± 0.03 6.82 ± 0.03 6.17 ± 0.03 6.03 ± 0.03 5.97 ± 0.06 5.83 ± 0.06

B 6.17 ± 0.03 6.27 ± 0.06 6.38 ± 0.03 6.48 ± 0.03 5.68 ± 0.03 5.58 ± 0.03 5.53 ± 0.03 5.47 ± 0.03

C 5.82 ± 0.03 6.02 ± 0.03 6.07 ± 0.06 6.12 ± 0.03 5.48 ± 0.06 5.38 ± 0.06 5.48 ± 0.03 5.33 ± 0.06


0

10

20

30

40

50

60

70

80

90

A B C

Mean p

art

icle

siz

e (

µm

)

0 kGy

5 kGy

7.5 kGy

10 kGy

0

1

2

3

4

5

6

7

8

A B C

pH

0 kGy

5 kGy

7.5 kGy

10 kGy

142


Table 8. Microbial contamination value of formulas before and after irradiation after 24 h of formulation and at 90th day of analyses.

C

o

d

e

Storage

conditions MC




5 7.5 10 5 7.5 10

A

Room

temperature

30 ± 2°C

B 3.55 × 104 3.15 × 103 0 0 2.14 × 107 3.31 × 105 0 0

MY 3.13 × 103 0 0 0 3.52 × 105 0 0 0

B B 4.09 × 104 4.09 × 103 0 0 2.55 × 107 3.88 × 105 0 0

MY 3.83 × 103 0 0 0 4.34 × 105 0 0 0

C B 4.67 × 104 5.08 × 103 0 0 3.02 × 107 4.39 × 105 0 0

MY 4.67 × 103 0 0 0 4.86 × 105 0 0 0

A

High

temperature

40 ± 2°C

B 4.15 × 104 3.92 × 103 0 0 2.50 × 107 4.08 × 105 0 0

MY 3.40 × 103 0 0 0 3.40 × 105 0 0 0

B B 4.58 × 104 4.82 × 103 0 0 2.93 × 107 4.82 × 105 0 0

MY 3.95 × 103 0 0 0 3.98 × 105 0 0 0

C B 5.51 × 104 5.96 × 103 0 0 3.42 × 107 5.59 × 105 0 0

MY 4.74 × 103 0 0 0 4.60 × 105 0 0 0

*MC: microbial contamination (cfu/g); B: bacteria; MY: mold-yeast; values were represented as mean ± standard deviation, n=3

Table 9. Rf values and densitogram peak areas of formula A before and after irradiation after 24 h of formulation and at 90th day of

analyses ( = 317 nm).

Peak Rf




5 7.5 10 5 7.5 10

1st 0.35 – 0.40 1777.8 2040.2 2564.5 3036.5 1237.8 1831.7 2492.7 3005.2

2nd 0.42 – 0.47 4527.4 5254.8 5854.1 6016.5 4045.3 4864.5 5802.3 5968.6

3rd 0.47 – 0.56 2854.1 2901 3061.1 4098.3 2451.3 2603.9 2998.2 4052.8

4th 0.62 – 0.72 16032.8 17858.1 19239.3 20587.5 14325.8 16622.6 19094.6 20462.4

5th 0.80 – 0.95 9231.9 12177.1 13625.1 14600.4 8101.9 11467.8 13394.8 14473.6

TLC-densitometry on formulas

Based on the densitogram 3D profiles

( = 317 nm) there were at least 5 peak areas

observed in formula A before and after irradiation

(Fig. 8).

Fig. 8. Densitogram 3D profiles of unirradiated and irradiated

formula A at 90 days of analyses ( = 317 nm).

Table 9 shows that there were an alteration

between unirradiated and irradiated peak areas.

The presence of peak area alterations allegedly due

to the effects of irradiation. Generally, it is shown

that irradiation dose up to 10 kGy caused increasing

of peak area. The increasing of peak area possible

due to the effects of irradiation that caused the

formation of new components. However, there were

also decreasing in peak areas, which is possibly

caused by degradation of the components into the

other components.

CONCLUSION

The use of gamma irradiation for

decontamination is advantageous for final cosmetic

products as well as raw materials. Sterilization is not

an obligation for cosmetic products. However,

they have to be protected from any contamination

or deterioration. The G. mangsotana pericarp

extract cream formulas with irradiation dose until

143


10 kGy presented acceptable physicochemical and

microbiological stabilities, for at least 90 days,

when the formulas were stored at room temperature

(30 ± 2°C) and at high temperature (40 ± 2°C).

Formula A which contained 1% of G. mangostana

pericarp extract with irradiation dose of 7.5 kGy is

the optimum formula from this research.

ACKNOWLEDGMENT

The authors wish to express our gratitude for

facilities and supports from Pancasila University and

Center for Application of Isotopes and Radiation

Technology, National Nuclear Energy Agency,

Indonesia.

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Documents

Formulation of Oil-in-Water Cream from Mangosteen